Prokaryotic expression of EV71 VP1 and initial evaluation of the biological activity

VernacularTitle:肠道病毒71型VP1的原核表达及生物活性初步研究
Author: Zhihui LI ; Nannan SONG ; Yingying YUE ; Peng LI ; Xuan JI ; Yinguang CAO ; Hong MENG
Publication Type:Journal Article
Keywords: Enterovirus 71; VP1; Prokaryotic expression; Biological activity
From: Chinese Journal of Microbiology and Immunology 2012;(11):972-976
CountryChina
Language:Chinese
Abstract: Objective To express EV71 VP1 in prokaryotic expression system,initially evaluate the ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody.Methods Construct the prokaryotic expression plasmid pET30a (+)-VP1.Induced expression in Transetta (DE3) with IPTG and identified by Western blot.After purified with HisBind Protein Purification Kit,its ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody were analyzed.Results Plasmid PET-30a(+)-VPI was constructed successfully and the objective protein was expressed effectively.The ELISA titer of the polyclonal antibody was 1:3200 while neutralizing titer was 1:16 and the recombination protein lost the ability of blocking EV71 infection.Conclusion The recombination protein can stimulate mice to produce antibodies and the polyclonal antibody shew neutralizing activity but the recombination protein lost the binding activity to receptors probably due to the wrong advanced structure.